Saturated and mono-unsaturated long-chain hydrocarbon profiles from Citrus unshiu juice sacs

Six cultivars of satsuma mandarin—Kawano Wase, Owari, Silverhill, Foley, Sugiyama and Nobilis—were examined for content of saturated and mono-unsaturated long-chain hydrocarbons in their juice sacs. Linear hydrocarbons accounted for more than 53% of the saturates and more than 87% of the monoenes. In the saturated fraction the major linear hydrocarbon was C₂₅ while in the monoene fraction C₂₅ and C₂₉ predominated. The ratios of linear/iso C₂₃ and C₂₅ saturates were greater than in other citrus previously investigated. Kawano Wase had profiles quite different from the other cultivars. Foley and Owari can be differentiated from the other cultivars by their linear monoene profiles. Long-chain hydrocarbon profiles intrinsic to the mandarin species, C. unshiu, were compared with profiles of non-mandarin species.     

Saturated mono-unsaturated long-chain hydrocarbon profiles of sweet oranges

Three varieties of midseason oranges, viz. Jaffa, Homosassa and Queen, were examined for their saturated and mono-unsaturated hydrocarbon composition in juice sacs. Hydrocarbons were isolated by lipid extraction of the juice sac powders followed by column, thin-layer and AgNO₃-TLC. After hydrogenation, the mono-unsaturated fraction and the saturated fraction were analyzed by GC. In the saturated fraction, the dominant linear hydrocarbon was C₂₃ while C₂₅ predominated in the monoene fraction. iso- And anteiso-branched hydrocarbons comprised between 53 and 63 % of the saturated fraction and only 20–26 % of the monoene fraction. Queen differed from Homosassa and Jaffa in the accumulation of higher percentages of saturated iso- and anteiso-branched hydrocarbons and conversely, showed lower percentages for these branched structures in the monoene fraction. Based on the total relative percentages of the three isomeric hydrocarbons, Homosassa could not be differentiated from Jaffa. The overall profiles for these two oranges, however. showed noticeable differences.     

Saturated and mono-unsaturated long-chain hydrocarbons from lemon juice sacs

Three varieties of lemon—Lisbon, Malta and Kusner—were examined for their content of juice sac saturated and mono-unsaturated long-chain hydrocarbons. The saturated fractions were 20 times the concentrations of the monoene fractions. The dominant linear hydrocarbon in the saturated fraction was C₂₅ while C₂₉ predominated in the monoene fraction. The saturated hydrocarbon profiles for Lisbon and Kusner were very similar to the profiles previously reported for Eureka lemon and Persian lime. The mono-unsaturated profiles were distinct for each of the three lemon varieties. In addition the lemon mono-unsaturated profiles were quite distinct from the hydrocarbon profiles previously reported for several other citrus species. In general the data support the elongation-decarboxylation mechanism for hydrocarbon synthesis proposed by Kolattukudy.     

Isoflavone,wax and triterpene constituents of Wyethia mollis

The hexane extract of Wyethia mollis contains the n-alkanes C₁₅-C₁₈, C₂₀-C₂₅, C₂₇ and C₂₉. Linoleic acid was the only detectable acidic component. A mass spectral analysis of the wax ester fraction indicated that it was a mixture of homologues, the saturated even-carbon acids n-C₁₆-C₃₀ esterfield with the saturated even-carbon alcohols n-C₁₈-C₂₆. The chloroform extract yielded the known isoflavones santal and 3′-O-methylorobol along with a new lanostane-type triterpene, 22,25-epoxy-lanosta-7:9(11)-dien-3-one. The wide distribution of n-alkanes and the decreased odd-even carbon ratio are consistent with the proposed primitive nature of this plant.     

Hydrocarbon production in Anacystis montana and Botryococcus braunii

The production of labelled aliphatic hydrocarbons in Anacystis montana and Botryococcus braunii has been studied using Na₂CO₃ [¹⁴C] as a carbon source. The major hydrocarbon produced by A. montana is pentadecane (ca 93%) accompanied by a pentadecene (ca 4%) and other hydrocarbons in the range C₁₃-C₁₇. Long chain (C₂₁-C ₃₃) hydrocarbons could not be detected in this organism. The variety of unsaturated hydrocarbons (C₂₅-C₃₁) previously reported in Botryococcus braunii is confirmed and contrasts with the synthesis of unsaturated C₁₇ hydrocarbons only, in axenic cultures prepared from single cell isolates of this colonial alga.     

Aliphatic Hydrocarbons of Cladosporium resinae Cultured on Glucose,Glutamic Acid,and Hydrocarbons

The carbon source markedly influenced the qualitative and quantitative composition of cellular hydrocarbons in Cladosporium resinae. Total lipid and hydrocarbon content was greater in cells grown on n-alkanes than in cells grown on glucose or glutamic acid. Glucose-grown cells contained a spectrum of aliphatic hydrocarbons from C₇ to C₃₆; pristane and n-hexadecane comprised 98% of the total. Cells grown on glutamic acid contained C₇ to C₂₃ hydrocarbons; n-tridecane, n-tetradecane, n-hexadecane, and pristane made up 74% of the total. n-Decane-grown cells yielded C₈ to C₃₂ compounds, and n-hexadecane (96%) was the major hydrocarbon. Cells grown on individual n-alkanes from C₁₁ to C₁₅ all contained C₁₁ to C₂₈ hydrocarbons, and cells grown on n-hexadecane contained C₁₁ to C₃₂ hydrocarbons. In n-undecane-grown cells, n-hexadecane and pristane made up 92% of the total, but in cells grown on C₁₂ to C₁₆ n-alkanes the major cellular hydrocarbon was the one on which the cells were grown. This suggests that cells cultured on n-alkanes of C₁₂ or longer accumulate n-alkanes prior to oxidizing them.     

Dependence of the hydrocarbon constituents of the leaf waxes of Khaya species on leaf age

The hydrocarbon constituents of the leaf waxes of eight species of Khaya were analysed for taxonomic purposes using GLC. The leaf waxes contained neither isoalkanes nor alkanes and the bulk of the n-alkanes were in the range of C₂₅ to C₃₃, odd-carbon number compounds predominating. It was found that the percentage composition of the n-alkane constituents of the leaf waxes varied with the age of leaves, young leaves having n-C₂₉ as the most abundant alkane, whereas older leaves had n- C₃₁.     

The solubility of hydrocarbon gases in lipid bilayers

The solubilities of 8 hydrocarbon gases in lipid bilayers have been determined at 25°C using a gas chromatographic technique. Results in 96% egg phosphatidylcholine, 4% phosphatidic acid bilayers expressed as Bunsen coefficients are: CH₄ 0.20; C₂H₆ 0.97; C₃H₈ 3.6; n-C₄H₁₀ 17; c-C₃H₄ 11; i-C₄H₁₀ 9.5; cis-butene 25; trans-butene 25. When 29 mol% cholesterol was incorporated with these phospholipids the Bunsen coefficient was reduced slightly. The observed solubilities are somewhat lower than those found in simple hydrocarbon solvents. The relative magnitudes of the solubilities of the gases in a given bilayer are closely related to the strength of their intermolecular forces.The solubility of butane was studied more extensively. No marked effects were observed when either surface charge or acyl chain saturation was increased, but the solubility in erythrocyte ghosts was much lower than that in lipid bilayers. The partition coefficient increased with decreasing temperature in egg lecithin and dimyristoylphosphatidylcholine. The entropy of solution in each case was more negative than that in isotropic non-polar solvents, and this accounts for the lower solubility in bilayers.The negative free energy of solution per CH₂ group for n-hydrocarbons is approximately twice that previously determined [11] for n-alcohols. This suggests that the concept of group contributions to solubility may not be universally applicable to lipid bilayers.     

Steroids,phthalyl esters and hydrocarbons from Balanites wilsoniana stem bark

IR assay for sapogenin of Balanites wilsoniana revealed 0.2% in the root wood, 0.7% in the root bark, 0.3% in the stem bark, 1.4% in the fatty seed and 0.6%. w/w in the leaf. The 25 α-epimers predominated in all parts except in the root wood. Six glucosideshaving diosgenin or yamogenin as aglycones were found and one was characterized diosgenin 3 β-d-glucopyranoside from IR, MS and NMR studies. Cholesterol, stigmasterol, sitosterol, 25 α-spirosta-3:5-diene and 25-β-spirosta-3:5-diene were also present. Free diosgenin and dienes were detected in appreciable quantities in the fat from the bark. The ‘unsaponifiable’ fraction of this fat contained phthalyl esters (mainly dioctyl and dibutyl) and saturated hydrocarbons C₁₀C₃₂ (with C₁₀C₂₀ predominating), together with the above mentioned steroids.     

Comparative Characterization of Extracellular and Intracellular Hydrocarbons of Clostridium pasteurianum

Extracellular and intracellular hydrocarbons produced by Clostridium pasteurianum VKM 1774 during cultivation on glucose-containing media in an argon atmosphere or in the presence of carbon dioxide and molecular hydrogen were analyzed by gas-liquid chromatography. Intracellular hydrocarbons were 50-55% (C₂₅-C₃₅) n-alkanes. Carbon dioxide and molecular hydrogen stimulated synthesis of extracellular hydrocarbons, which comprised 90-95% (C₁₁-C₂₄) n-alkanes.     

Hydrocarbons,phenols and sterols of the testa and pigment strand in the grain of Hordeum distichon

Material from the testa of decorticated barley grains contained hydrocarbons, esters, triglycerides, free sterols, 5-n-alkylresorcinols, and traces of free alcohols, carbonyl compounds, and various polar, acidic materials. The hydrocarbon fraction was mainly a series of n-alkanes, extending at least from C₁₁ to C₃₆, in which the C₂₉ and C₃₁ components were prominent. Two minor series of alkanes were also present. Sometimes a trace of an unsaturated hydrocarbon was detected. The ester fraction contained sterols and alkanols esterified by fatty acids, which differed in relative amounts from the fatty acids found in the triglycerides. The triglycerides were thought to have leached from within the grain. At least five free sterols were present, including sitosterol and campesterol. The 5-n-alkylresorcinols were at least twelve members of a homologous series, of which four, C₂₅, C₂₇, C₂₉, and C₃₁, made 98% of the total. Members of the series with even numbers of carbon atoms were also present. It is suggested that they are partly responsible for excluding microorganisms from the interior of the grain. The testa membrane, with the associated pigment strand, contained an estolide of fatty acids and various hydroxyacids, a polysaccharide component, and uncharacterized material.     

Comparative alkane chemistry of Parthenium argentatum (guayule) and some F1 hybrids

The leaf alkanes of Parthenium argentatum (guayule), P. tomentosum var. stramonium, P. fruticosum var. trilobatum, and the first filial (F₁) generations obtained from crosses with guayule were investigated by GC and mass spectrometry and shown to be useful in chemotaxonomic studies. The identified n-alkanes ranged from C₁₉ to C₄₀ with either n-C₂₉ or n-C₃₁ as the main component. The alkane chemistry of guayule with n-C₃₁ being the main component predominated in most of the F₁ hybrids. The presence of iso-branched alkanes (C₂₇, C₂₉, C₃₁) in P. tomentosum and its hybrids could be detected by GC/MS. These preliminary investigations indicate that epicuticular wax alkanes can be useful in inheritance studies of guayule and its hybrids.     

Hydrocarbons,sterols and tocopherols in the seeds of six Adansonia species

The composition of the unsaponifiable matter of the lipids of six Adansonia species (A. grandidieri, A. za, A. fony, A. madagascariensis, A. digitata and A. suarezensis) was investigated. The total unsaponifiable content, its general composition and the identity of the components of the hydrocarbon, sterol and tocopherol fractions are presented. The unsaponifiable content in oil ranges from 0.4 to 1.1% (hexane method) and from 0.6 to 2.2% (diethyl ether method). In two species (A. grandidieri and A. suarezensis) the major components are 4-demethylsterols (23–42%) tocopherols (37-10%) and hydrocarbons (15–17%). In both species examined, eight 4-demethylsterols occur in the sterol fraction with sitosterol (81–88%) being predominant. Among the four tocopherols present, γ-tocopherol (68–98%) is the major compound. Each Adansonia species shows a characteristic gas liquid chromatography pattern for the hydrocarbon fraction. Squalene is the major component for five species (40–75%). Iso-, anteiso- and other branched hydrocarbons were not identified but were present in small amounts in comparison with n-alkanes. The dominance of odd- over even-carbon number chain length of n-alkanes was not observed in any species. The results show that C₂₂, C₂₅, C₂₆, C₂₇, C₂₈ and C₂₉ are the most frequent major constituents.     

Organic chemistry of Protosalvinia (Foerstia) from the Chattanooga and New Albany Shales

Vertical samplings of Protosalvinia, a thalloid Upper Devonian alga from the Chattanooga and New Albany Shales, are chemically analyzed and correlated with the organic chemical constituents isolated from associated shale matrices. Normal, saturated acids (n-C₈ to n-C₃₆n-paraffins (C₁₀ to C₃₆), showing an odd carbon-number preference, branched-chain alkanes, and vanadyl porphyrins isolated from Protosalvinia vary in their concentrations with depth of burial and with the dominant associated morphology of Protosalvinia, i.e., P. arnoldii, P. ravenna, P. furcata. Organic constituents of shales, in general, reflect those detected in thalli; relative concentrations, molecular diversity, carbon chain-lengths and maxima of compounds extracted from both shale and fossil material are similar. Pristane, phytane and porphyrins are probably derived from a chlorophyllous organism, while δ¹³C data corroborate a photosynthetic system operating in the primary biosynthesis of shale geochemistry. Crude-oil extracts of Protosalvinia-rich strata contain higher alkane and lower aromatic hydrocarbon concentrations than those of an average crude oil. Chemical variations among forms of Protosalvinia suggest biochemical differences in original plant composition rather than diagenetic transitions; field observations of morphological trends seen in vertical samplings may be used in crude extrapolations of the organic chemical compositions of shale strata.     

Alkadiene- and botryococcene-producing races of wild strains of Botryococcus braunii

Samples of the green colonial alga Botryococcus braunii, collected from various localities, were grown in the laboratory and examined for their hydrocarbon content and morphology. Although few differences appeared between the ultrastructures of the samples, the nature of their hydrocarbons, which remains unchanged at any stage of growth, allows the distinction of two physiological races viz algae producing odd-numbered unbranched alkadienes and trienes (C₂₅C₃₁) (the A race) and those producing polymethylated triterpenes C_nH_2n-₁₀ (C₃₀C₃₇), the botryococcenes (the B race). In laboratory culture, the hydrocarbon content of these new strains is very high, from 30 to 60% of the dry biomass. For the two races the greatest hydrocarbon productivity takes place during the active growth phase. The important variability observed in botryococcene distribution could originate both from genetic and environmental factors.     

Incorporation of labelled dietary n-alkanes into cuticular lipids of the grasshopper Melanoplus sanguinipes

Dietary hydrocarbons are incorporated into cuticular lipids of the grasshopper Melanoplus sanguinipes. Dietary secondary alcohols and ketones, however, are not incorporated into the cuticular lipids. In typical experiments from 8 to 28 per cent of the fed labeled n-alkanes are recovered in the cuticular lipids. Most of the radioactivity recovered from feeding the C₂₃ n-alkane and a significant amount from the C₂₅ was found as a secondary alcohol in the form of a wax ester. The C₂₉ and C₃₁ n-alkanes were recovered primarily unchanged as the n-alkane. Eighty-five per cent of injected acetate incorporated into the hydrocarbon fraction is in the branched hydrocarbons. These results show that the insect synthesizes its branched hydrocarbons, whereas a large part of the normal hydrocarbons can be dietary.     

Tests Whether a Head to Head Condensation Mechanism Occurs in the Biosynthesis of n-Hentriacontane, the Paraffin of Spinach and Pea Leaves

Isobutyrate-1-¹⁴C and l-isoleucine-U-¹⁴C fed through the petiole labeled the surface lipids of broccoli leaves, but the incorporation was much less than from straight chain precursors. Not more than one-third of the ¹⁴C incorporated into the surface lipids was found in the C₂₉ paraffin and derivatives, whereas more than two-thirds of the ¹⁴C from straight chain precursors are usually found in these compounds. The small amount of ¹⁴C incorporated into the paraffin fraction was found in the n-C₂₉ paraffin rather than branched paraffins showing that the ¹⁴C in the paraffin must have come from degradation products. Radio gas-liquid chromatography of the saturated fatty acids showed that, in addition to the n-C₁₆ acid which was formed from both branched precursors, isoleucine-U-¹⁴C gave rise to branched C₁₅, C₁₇, and C₁₉ fatty acids, and isobutyrate-1-¹⁴C gave rise to branched C₁₆ and C₁₈ acids. Thus the reason for the failure of broccoli leaf to incorporate branched precursors into branched paraffins is not the unavailability of branched fatty acids, but the absolute specificity of the system that synthesizes paraffins, probably the elongation-decar-boxylation enzyme complex. Consistent with this view, no labeled branched fatty acids longer than C₁₉ could be found in the broccoli leaf. The branched fatty acids were also found in the surface lipids indicating that the epidermal layer of cells did have access to branched chains. Thus the paraffin synthesizing enzyme system is specific for straight chains in broccoli, but the fatty acid synthetase is not.     

Hydrocarbons and fatty acids of Lycopodium

The analysis of fatty acids and hydrocarbons in the sporophytes of three Lycopodium species has revealed a characteristic distribution of C₁₆ and C₁₈ acids. The hydrocarbon fraction of the lipids contain a homologous series of monounsaturated alkenes in the C₁₇C₃₀ range with an even to odd preference. Maxima at both C₁₇ and C₂₇ among the n-alkanes reveals similarities both to the distribution of hydrocarbons in other plant groups. The production of spores and their inclusion with one sporophyte does not alter the fatty acid pattern but does decrease the alkene concentration and modifies the alkane distribution, shifting both maxima. The presence of pristane and phytane in all specimens, the dual maxima of alkanes and slight odd to even preference of alkanes is noteworthy in that these characteristics are possessed by geological deposits derived from Lycopodium ancestors.     

Production of Proteinase on Noncarbohydrate Carbon Sources by Pseudomonas aeruginosa

Proteinase production by Pseudomonas aeruginosa was studied in medium containing noncarbohydrate materials, especially various hydrocarbons, as the sole carbon source. On heavy oil, kerosene, n-paraffinic hydrocarbon of C₁₂, C₁₄, or C₁₆, and propylene glycol, the bacteria grew well and high protinase production was observed. However, production on paraffinic hydrocarbon differed remarkably with strains of varied origins. The elastase-positive strain, IFO 3455, showed abundant growth and high proteinase production on medium containing a paraffin of C₁₂, C₁₄, or C₁₆, whereas the elastase-negative strain, IFO 3080, showed little growth on the same medium. Neither elastase-positive nor elastase-negative strains, however, utilized n-paraffins of C₅ to C₁₀, or various aromatic hydrocarbons such as benzene, naphthalene, phenanthrene, and anthracene. The proteinases produced on the noncarbohydrate medium were identical with those produced in glucose medium.     

A Single Column Packing for The Gas-Liquid Chromatographic Separation of Various Biologically Important Compounds

The use of a single, commercially available column packing, Tabsorb_R, is described for the g. l. c. separation of a large number of different compounds. The resolution of the homologous members of the following series of compounds was achieved: (1) saturated fatty acids (C₁-C₁₈), (2) normal aliphatic saturated dlcarboxylic acids (C₂-C₁₄), (3) normal aliphatic saturated alcohols (C₁-C₂₄), (4) normal aliphatic saturated amines (C₁C₁₂), (5) the common amino acids except arginlne, histidlne and cysteine, (6) aliphatic hydrocarbons (C₁₀-C₂₀) and (7) monosaccharides. It should be noted that twenty-two monosaccharides including three hexosamines and two anhydrohexoses, could be resolved as aldltol acetates In a single run. In addition, galacturonic, glucuronic and lduronlc acids could be separated from one another as their 1, 4-lactones. The resolution achieved in these series of compounds was found to be consistent and highly reproducible. It is of further interest that certain Isomers of the higher fatty acids and hydrocarbons with one double bond could also be separated from the normal and saturated compounds, respectively. The applicability of “Tabsorb” for the g. l. c. separation, although noted above to be considerably broad, is by far not yet exhausted. These procedures which form the basis for the quantitative determinations of the various compounds studied as demonstrated by analysis of glycopeptldes for neutral hexoses and proteins for the amino acids, can readily be adapted to preparative methods. From the biochemical point of view “Tab-sorb” is an extremely versatile column packing in that it can be used for the identification of many of the common building blocks of natural products.